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 REPPCR 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

comparative study on some cardiopulmonary effects, anesthesia quality, and recovery time of isoflurane vs. propofol in domestic pigeons [Columba livia domesticus]

It is commonly acknowledged that the most safe and method of choice anesthesia in birds is inhalation anesthesia but in some clinical situations, such as tracheal resection, injectable anesthetic agents are the only choice of surgeons regardless of whether or not an anesthesia machine is available. This study aimed to compare the quality of anesthesia and recovery time of isoflurane and propofol in domestic pigeons. Twenty pigeons [Columba livia domesticus], weighing 302.5 +/- 37.95g [Mean +/- SD] were randomly allocated to two groups often. One group was anesthetized by isoflurane [Iso-group], and the anesthesia lasted for 30 minutes. The other group received 14 mg/kg of propofol [1%] at constant rate [CRI] through basilica [wing] vein catheter to induce anesthesia [Pro-group]. 1.33 mg/kg per min of propofol was infused to keep pigeons anesthetized for 30 minutes, using an injection pump. Temperature, heart rate, respiratory rate, and percentage of oxygen saturation of hemoglobin [SpO2%] were recorded in all three phases including before induction of anesthesia, during anesthesia at minutes 1, 3, 5, 10, 15, 20, 25 and 30, and after recovery time in both groups. Anesthesia caused significant effects on respiratory rate, heart rate, and SpO2% [p<0.05]. Recovery times in both groups were significantly different [longer in propofol group]. Our findings revealed that the pigeons anesthetized with isoflurane have a soft and fast anesthesia; however, the pigeons were anesthetized with propofol, had a rough induction that was not uniform for all pigeons. Isoflurane showed that it is safer than propofol to anesthetize pigeons

Identification of Coxiella burnetii by touch-down PCR assay in unpasteurized milk and dairy products in north - east of Iran

Coxiella burnetii is the causative agent of the zoonotic disease Q fever, and ruminants being considered as the main source for human infection. Although the main route of infection in human is inhalation of contaminated aerosols, oral transmission by contaminated raw milk or unpasteurized dairy products is also a possible route of infection. Raw milk or dairy products produced from unpasteurized milk may contain virulent C. burnetii. This study aimed to determine the contamination rate of milk and unpasteurized dairy products with C. burnetii. Touch-down PCR was used to examine the presence of C. burnetii on 147 dairy product samples collected from local traditional and commercial markets in Mashhad-Khorasan Razavi province- Iran. 2 of 28 [7.14%] cheese samples, 2 of 26 [7.69%] yoghurt samples, 8 of 23 [34.78%] sheep milk samples, and 2 of 60 [3.33%] cow milk samples were found to be positive for C. burnetii DNA. However, 10 goat milk samples were found to be negative. The results of this study indicate that the clinically healthy dairy livestock and their dairy products are important sources of C. burnetii infection

Intestinal colonization of different Brachyspira spp. in laying hens

Avian intestinal spirochetosis [AIS] is caused by spiral-shaped Gram-negative Brachyspira spp. in poultry. It is known as a cause of diarrhea, low egg production, and increased occurrence of dirty eggs in layer hens. In this study, the presence of some Brachyspira spp. was investigated in laying hens. A total of 100 cloacal swab samples were individually collected from 20 laying hen flocks showing fecal egg staining in northeast of Iran. Using culture and morphologic examination, 41 samples [41%] from 20 flocks were positive; however, by using genus-specific PCR, only 37 [37%] samples were confirmed as Brachyspira spp. Using species-specific primers, single colonization was identified in 18 samples associated with B. pilosicoli [48.6%], while single colonization with B. intermedia was found in only two samples [5.4%]. Simultaneous colonization by B. intermedia and B. murdochii was detected in 3 samples [8.1%]. B. pilosicoli was the most prevalent species in concurrent colonization in 11 cases [29.7%]. Finally, cocolonization by B. intermedia and B. innocens was identified in 3 samples [8.1%]. The results of this study showed the colonization of different species of Brachyspira with dominance of B. pilosicoli in layer hens

Isolation, antimicrobial susceptibility and mecA gene analysis of methicillin-resistant Staphylococcus aureus in Iranian white cheeses

The objective of this study was to determine the occurrence of antimicrobial resistance among Staphylococcus aureus and to estimate the presence of methicillin-resistance in S. aureus [MRSA] isolates obtained by culture and polymerase chain reaction [PCR] methods. For this purposes, 100 Iranian white and feta cheese samples collected from different suppliers were initially evaluated for the occurrence of S. aureus by culturing methods. The obtained isolates were subjected to disc diffusion antimicrobial susceptibility tests and a PCR method to detect the mecA gene. Out of the 100 cheese samples examined, 25 [25%] samples were contaminated with S. aureus with a mean of 5.74 +/- 5.67 log cfu/g. Out of the 25 isolates, 23 [92%] were found to be resistant to at least one antibiotic or more, tested by a disk diffusion method. The highest rate of antibiotic resistance was observed to penicillin G [92%] followed by ampicillin [73%] and cloxacillin [68%]. None of the isolates was resistant to gentamycin and vancomycin. Eight [34.78%] of the 23 S. aureus isolates were genotypically confirmed as MRSA. The results indicate that the presence of antimicrobial resistant strains of S. aureus in Iranian cheese samples constitute a potential risk for human health. This calls for better control of the spread of antimicrobial resistant strains as well as cheese contamination sources

Molecular characterization of avian adenoviruses in Iranian broiler flocks

This study was conducted to molecularly detect avian adenoviruses in broiler flocks showing liver lesions and respiratory syndrome in northeast Iran. In total, 60 tissue samples were collected from broiler farms with liver lesions, respiratory syndrome and also from clinically healthy flocks. Six samples were positive; however, three samples were selected for molecular studies. PCR products were sequenced to confirm the identity of avian adenoviruse. Based on the sequence analysis of the L1 region of the hexon gene, the NRB/FAV/4 should be classified as FAdV 8b strain and two other isolates - NRB/FAV/1 and NRB/FAV/5 - classified in cluster of the FAdV 2 and 11. As far as we know, this preliminary investigation is the only documented study to confirm the presence of avian adenoviruses in broiler flocks in Iran

Sequence analysis of the VP1 gene in three very virulent infectious Iranian bursal disease virus strains

Infectious bursal disease [IBD] is a highly contagious disease of chickens caused by the infectious bursal disease virus [IBDV]. This study was conducted to characterize three IBDV strains from Iran. A reverse transcriptase-polymerase chain reaction [RT-PCR] procedure was used to amplify a 715-bp fragment of the VP1 gene from IBDV strains. Amplified VP1 fragments of the three Iranian IBDV strains were sequenced and compared with published sequences of IBDV strains from around the world, and their phylogenetic relationships were analyzed. Alignment of IBDV strains revealed 23 nucleotide differences between vvIBDV [except for IL and PT] and other non-vvIBDV strains. Two nucleotide positions, 863G and 1023A, were specific as JRMP07IR and JRMP14IR strains. All vvIBDVs differed [except for IL and PT strains] from non-vvIBDVs at aa [amino acids] positions 242E and 287A. In the three Iranian IBDV strains, aa positions 251R in both JRMP07IR and JRMP14IR, and 360L in JRMP14IR differed from those of other vvIBDVs. In phylogenetic analyses, all three Iranian strains clustered together with vvIBDVs. One Iranian strain, JRMP30IR, was more closely related to two European strains [HOL and UK661] and two south-east Asian strains [OKYM and ZJ2000]. However, the other two Iranian strains, JRMP07IR and JRMP14IR, were closer to two Turkish strains [OA/G1 and OE/G2] and a Malaysian strain [UPM94]. Further comprehensive investigations will provide researchers a better knowledge on the distribution, variability, and phylogenetic relationships of different IBDVs isolated in Iranandother parts of the world. Key words: Infectious bursal disease virus, Very virulent strains, VP1, Chicken, Iran

PCR-based detection of cow and goat milk in sheep milk and dairy products marketed in Mashhad city of Iran

The extensive consumption of milk and dairy products makes these foodstuffs targets for potential adulteration with financial gains for unscrupulous producers. The aim of this study was using PCR assay to detect cow milk in labeled sheep milk, sheep yoghurt, and Lighvan cheese [a traditional ripened cheese produced from sheep's milk]. The assay utilized primers targeting the mitochondrial 12s and 16s rRNA gene. In this study, 35 samples of sheep milk, 35 samples of sheep yoghurt, and 35 samples of Lighvan cheese were purchased from different supermarkets in Mashhad city with different batch numbers. The results showed only 21 out of 105 [20%] samples contained pure sheep milk. Undeclared presence of cow and goat milk was detected in 33[31.5%] and 68[65%] of the 105 samples, respectively. It seems the PCR based analytical method is an applicable technique to monitor adulteration in dairy products

Mixed cutaneous round cells tumor in a cock [Gallus domesticus]: a case report

Cutaneous round cell tumors have been classified as mast cell tumor [MCT], histiocytoma [HCT], lymphosarcoma, undifferentiated round cell tumors and occasionally rhabdomyosarcoma in veterinary medicine. An adult cock [Gallus domesticus] showing a large solitary integument mass raised on dorso bilateral of cervical part, extending to intercapsular and cranial mid part of the back was referred to the birds clinic of the faculty of veterinary medicine at the university of Tehran. Microscopic examination revealed sheeted cells with large, round to oval nuclei with each one containing one or more prominent nucleoli with scant cytoplasm. The myofibrils of the neck were degenerated by aggressive tumor cells. The condition was differentiated from other round cell tumors by electron microscope, histochemical staining, as well as the application of a large panel of antibodies. Polymerase chain reaction failed to confirm the involvement of both Marek's disease virus and avian leukosis virus subgroup-J. It was concluded that the tumor cells were consistent with both B-cell lymphocytes and histiocytes that unusually covered the entire dermal layer of dorsal neck skin. This unusual cutaneous lymphoma was named as lymphoblastic histiocytoma

Psittacine beak and feather disease in Iran, molecular and histopathologic detection

Psittacine beak and feather disease [PBFD] is a major viral disease in wild and captive psittaciformes all around the world. The disease was suspected in a 7 years old lesser sulphur-crested cockatoos [Cacatua sulphured] with a minor feather loss at the back of neck and head. The bird was comprehensively examined by macroscopic pathology, histopathology and polymerase chain reaction [PCR]. Marked intracellular edema of the keratinocytes and necrosis were evident in histopathological observation of dystrophic feather follicles. Numerous macrophages with cytoplasmic inclusions [botryoid] and Prevasculitis were also present in the dermis. Histopathologically, the feather lesions and inclusions were typical of PBFD. The presence of psittacine beak and feather disease virus [BFDV] DNA was confirmed by PCR. This is the first documented report of the occurrence of the PBFD in Iran

[Rapid differentiation between very virulent and classical infectious bursal disease viruses isolated in Iran by RT-PCR/REA]

This study was conducted to characterize infectious bursal disease virus [IBDV] isolates collected from different parts of Iran during 2005-2006. Pooled bursal samples from 49 broiler and layer pullet flocks suspected to IBD infection were collected and processed. A reverse transcriptase-polymerase chain reaction [RT PCR] procedure was used to amplify a VP2 gene fragment [743 bp] from IBDV field isolates. Amplified VP2 fragments were further characterized by two restriction enzymes, BspMI and Sad. From 49 field samples, 37 [75.5%] samples were IBDV positive by RT PCR. Digestion with two restriction enzymes, BspMI and SacI, showed patterns compatible with very virulent IBDV and classical IBDV strains in 34 [91.9%] and 3 [8.1%] IBDV-positive samples, respectively. The restriction enzyme analysis of this study was comparable to that of other isolates and reference strains with available nucleotide sequence data in the GenBank. The procedure followed in this study is a useful method to rapidly differentiate the very virulent IBDV and classical IBDV isolates

Cholangiocarcinoma in a broiler chicken

Twenty carcasses from a small flock of 5000 four-weeks old broiler chickens were submitted to a poultry disease diagnostic clinic in Tehran. At necropsy, 19 carcasses showed the typical lesions of colibacillosis such as pericarditis, prihepatitis and airsaculitis. One case did not show any gross lesion of colibacillosis or chronic respiratory disease [CRD] but the liver had an abnormal size containing a cystic part at the base of right lobe filled with a whitish fluid. In bacteriological culture, proteus was isolated from liver. Histopathological examination revealed multicentric bile duct hyperplasia and cholangiocarcinoma in the liver. Neoplastic cells effaced hepatocellular architecture. Histological examination of the neoplastic areas in the liver revealed cholangiocarcinoma [bile duct carcinoma]. This appears to be the first reported case of cholangiocarcinoma in birds in Iran

Case report: Lipoma in a budgerigar [Melopisittacus undulates]

To determine the identity and the cause ofmasses infiltrated in the most part of the body of abudgerigar. Case report. A pair of budgerigars [Melopisittacus undulates], living together and using the same diet for 4years, were presented to the Birds Clinic of PoultryDiseases Section, Department of Clinical Sciences, Facultyof Veterinary Medicine, University of Tehran. Clinical examination, radiography, necropsy,gross and histopathological observations. The male bird had no obvious health problem butthe female one demonstrated dyspnea, abnormal growth ofthe beak and the presence of hard and elevated masses inthe thoracic and abdominal area. Radiography revealed thepresence of masses infiltrated in the most part of the bodyand in particular at the opening of the thoracic andabdominal cavities. Due to critical condition of the femalebird and at its owner consent, the bird was euthanatized andnecropsied. Yellow-colored masses with a relatively hardstructure at the opening of the thoracic and abdominalcavities and soft fat tissues in the other parts of the body [including underarm and inguinal area] were observed innecropsy. Histological examination of the masses wasdiagnosed as lipoma

[ cryptosporidum muris like parasite in large ruminants in various parts of Iran]

Faecal samples from 3470 cattle, buffalos, camels and common mouse were collected from various farms located in and around Tehran, Isfahan, Shiraz, Tabriz, Mashad, Zahedan and Ahwaz. They were examined for the presence of C. muris - like oocysts using modified Ziehl -Neelsen technique staining. Within this study, it was observed that whereas some farms were completely negative others showed a varying degree of positive cases. For instance, in one of the farm tested at Isfahan 12% of cows were positive for C. muris - like. The presence of C. muris - like in mice circumstantially suggests the possibility of mice transmitting the infection to cattle but it is not known whether cattle can actually become infected with C. muris - like oocysts from mice. The present article might be the first report en the presence. muris - like organism in large ruminants and brown mice in various parts of Iran